Human Kallikrein 4 ELISA Kit

From Antibodies.com, this sandwich ELISA enables the in vitro quantitative determination of human Kallikrein 4 in serum, plasma or other biological fluids, with reactivity confirmed for human samples [S1]. The assay uses a pre-coated 96-well microplate (12 x 8 well strips) and follows the sandwich enzyme immunoassay technique [S1]. An antibody specific for Kallikrein 4 is immobilised on the plate; standards and test samples are added, and the Kallikrein 4 present in each sample is captured by the coated antibody [S1]. Following incubation and washing, a Biotinylated Anti-Kallikrein 4 Antibody is added, which binds the captured Kallikrein 4, followed by HRP-Avidin conjugate [S1]. A TMB substrate solution visualises the HRP enzymatic reaction, producing a blue-coloured product that changes to yellow upon addition of acidic stop solution [S1]. The density of yellow is proportional to the amount of Kallikrein 4 captured in each well, and the concentration is calculated by reading the O.D. absorbance at 450 nm in a colourimetric microplate reader against a standard curve [S1]. Total assay time is 4h 30m [S1].

Stated sensitivity is <0.097 ng/ml and the assay range is 0.312-20 ng/ml [S1]. Reported spike-and-recovery performance in serum was 80%-102% (average 91%, n=5); in EDTA plasma, 81%-100% (average 90%, n=5); and in heparin plasma, 80%-89% (average 84%, n=5) [S1]. Dilution linearity at 1:2, 1:4, 1:8 and 1:16 in serum was 87-91%, 87-107%, 74-101% and 92-97% respectively; in EDTA plasma, 90-105%, 84-101%, 90-101% and 79-108%; and in heparin plasma, 84-95%, 92-105%, 82-105% and 89-91% [S1]. Kit components include the pre-coated 96 well microplate (12 x 8 well strips), lyophilised standard, detection solutions A and B, 30X wash buffer, sample dilution buffer, TMB substrate, stop solution and plate sealers, stored at +4°C or -20°C as instructed [S1].